Scanning the active center of formolase to identify key residues for enhanced C1 to C3 bioconversion.

BACKGROUND: Formolase (FLS) is a computationally designed enzyme that catalyzes the carboligation of two or three C1 formaldehyde molecules into C2 glycolaldehyde or C3 dihydroxyacetone (DHA). FLS lays the foundation for several artificial carbon fixation and valorization pathways, such as the artificial starch anabolic pathway. However, the application of FLS is limited by its low catalytic activity and product promiscuity. FINDINGS: FLS, designed and engineered based on benzoylformate decarboxylase from Pseudomonas putida, was selected as a candidate for modification. To evaluate its catalytic activity, 25 residues located within an 8 Å distance from the active center were screened using single-point saturation mutagenesis. A screening approach based on the color reaction of the DHA product was applied to identify the desired FLS variants. After screening approximately 5,000 variants (approximately 200 transformants per site), several amino acid sites that were not identified by directed evolution were found to improve DHA formation. The serine-to-phenylalanine substitution at position 236 improved the activity towards DHA formation by 7.6-fold. Molecular dynamics simulations suggested that the mutation increased local hydrophobicity at the active site, predisposing the cofactor-C2 intermediate to nucleophilic attack by the third formaldehyde molecule for subsequent DHA generation. CONCLUSIONS: This study provides improved FLS variants and valuable information into the influence of residues adjacent to the active center affecting catalytic efficiency, which can guide the rational engineering or directed evolution of FLS to optimize its performance in artificial carbon fixation and valorization.

New eremophilane-type sesquiterpenes from Synotis solidaginea.

Eleven new highly oxygenated eremophilane-type sesquiterpenoids were isolated from the whole plant of Synotis solidaginea, including two pairs of C-8 S/R epimers. The structures of the new compounds were elucidated on the basis of detailed spectroscopic analysis and the absolute configurations of 1 and 9 were confirmed by single-crystal X-ray crystallography using Cu Kα radiation. All the isolates were tested for the inhibition of LPS-stimulated NO production in macrophage-like mouse monocytic leukemia RAW264.7 cells. Compound 1 exhibited weak inhibitory effects with an IC50 of 71.2 µM.

Development and Application of a Comprehensive Nontargeted Screening Strategy for Aristolochic Acid Analogues.

Aristolochic acid analogs (AAAs) are naturally occurring carcinogenic and toxic compounds that pose a safety threat to pharmaceuticals and the environment. It is challenging to screen AAAs due to their lack of characteristic mass spectral fragmentation and their presence of structural diversity. A comprehensive nontargeted screening strategy was proposed by taking into account diverse factors and incorporating various self-developed techniques, and a Python3-based toolkit called AAAs_finder was developed for its implementation. The main procedures consist of virtual structure and ultraviolet and visible (UV) spectra database creation, exact mass and UV spectra-based suspect data extraction, tandem mass spectra (MS/MS) anthropomorphic interpretation, and multicondition retention time (RT) prediction-based candidate structures ranking. To initially assess screening feasibility, eight hypothetical unknown samples were subjected to nontargeted screening using the AAAs_finder toolkit and two other advanced tools. The results showed that the former successfully identified all, while the latter two only managed to identify two and three, respectively, indicating that our strategy was more feasible. After that, the strategy was carefully evaluated for false positives and false negatives, instrument dependence, reproducibility, and sensitivity. After the evaluation, the strategy was successfully applied to the screening of AAAs in real samples, such as herbal medicine, spiked soil, and water. Overall, this study proposed a nontargeted screening strategy and toolkit independent of characteristic mass spectral fragmentation and able to overcome challenges posed by structural diversity for the AAAs screening, which is also valuable for other classes of compounds.

Iodine Intake Trends in United States Girls and Women between 2011 and 2020.

BACKGROUND: Usual intakes of iodine in United States girls and women, including pregnant and lactating women have not been adequately studied. Adequate intake of iodine is critical for neurodevelopment of girls, thyroid functions, and reproductive health of women. OBJECTIVES: This study aimed to examine the adequacy and trends of iodine intake of United States girls and women between 2011 and 2020. METHODS: We mapped the sources of United States girls and women's iodine intake from the 29 food groups between 2011 and 2020 using United States Department of Agriculture's iodine data release 2. The total food intakes from 2 d of dietary recall of the United States National Health and Nutritional Examination Survey and estimated iodine concentrations of the food groups were used to calculate the usual iodine intakes of female participants. Trends of usual intakes, urinary iodine concentrations (UIC), and estimated intake adequacy were calculated. RESULTS: Median usual intakes of iodine estimated from diet and supplements and UIC of United States girls and nonpregnant, nonlactating women declined between 2011 and 2020 in all 3 age groups: ≤14 y, 15-49 y old, and ≥50 y. Median usual intakes of iodine for pregnant and lactating United States women declined as well. Inadequacy levels of usual iodine intake were 9.9% for nonpregnant, nonlactating women of reproductive age 15-49 y old, 40.3% for lactating, and 10.2% for pregnant women in the 2017-2020 period. Intake insufficiencies estimated from UIC were 48.8%, 63.2%, and 31.3% for nonpregnant, nonlactating women of reproductive age 15-49 y old, pregnant and lactating women, respectively, in the 2017-2020 period. A significant decline in milk consumption might be one of the major contributors to the dietary iodine decline in United States women. CONCLUSIONS: Iodine intake of United States girls and women were on the decline between 2011 and 2020 and the increased inadequacy of iodine intake deserves public health attention.

Genome-Wide Identified MADS-Box Genes in Prunus campanulata 'Plena' and Theirs Roles in Double-Flower Development.

The MADS-box gene family plays key roles in flower induction, floral initiation, and floral morphogenesis in flowering plants. To understand their functions in the double-flower formation of Prunus campanulata 'Plena' (hereafter referred to as PCP), which is an excellent flowering cherry cultivar, we performed genome-wide identification of the MADS-box gene family. In this study, 71 MADS-box genes were identified and grouped into the Mα, Mß, Mγ and MIKC subfamilies according to their structures and phylogenetic relationships. All 71 MADS-box genes were located on eight chromosomes of PCP. Analysis of the cis-acting elements in the promoter region of MADS-box genes indicated that they were associated mainly with auxin, abscisic acid, gibberellin, MeJA (methyl jasmonate), and salicylic acid responsiveness, which may be involved in floral development and differentiation. By observing the floral organ phenotype, we found that the double-flower phenotype of PCP originated from petaloid stamens. The analysis of MIKC-type MADS-box genes in PCP vegetative and floral organs by qRT-PCR revealed six upregulated genes involved in petal development and three downregulated genes participating in stamen identity. Comparative analysis of petaloid stamens and normal stamens also indicated that the expression level of the AG gene (PcMADS40) was significantly reduced. Thus, we speculated that these upregulated and downregulated genes, especially PcMADS40, may lead to petaloid stamen formation and thus double flowers. This study lays a theoretical foundation for MADS-box gene identification and classification and studying the molecular mechanism underlying double flowers in other ornamental plants.

Rapid recognition and targeted isolation of potential anti-breast cancer xanthones in Hypericum bellum Li by "seed" mass spectra-based molecular networking and in silico MS/MS fragmentation.

INSTRUCTION: Hypericum bellum Li is rich in xanthones with various bioactivities, especially in anti-breast cancer. While the scarcity of mass spectral data of xanthones in Global Natural Products Social Molecular Networking (GNPS) libraries have challenged the rapid recognition of xanthones with similar structures. OBJECTIVE: This study is aimed to enhance the molecular networking (MN)-based dereplication and visualisation ability of potential anti-breast cancer xanthones from H. bellum to overcome the scarcity of xanthones mass spectral data in GNPS libraries. Separating and purifying the MN-screening bioactive xanthones to verify the practicality and accuracy of this rapid recognition strategy. METHODOLOGY: A combined strategy of "seed" mass spectra-based MN, in silico annotation tools, substructure identification tools, reverse molecular docking, ADMET screening, molecular dynamics (MDs) simulation experiments, and an MN-oriented separation procedure was first introduced to facilitate the rapid recognition and targeted isolation of potential anti-breast cancer xanthones in H. bellum. RESULTS: A total of 41 xanthones could only be tentatively identified. Among them, eight xanthones were screened to have potential anti-breast cancer activities, and six xanthones that were initially reported in H. bellum were obtained and verified to have good binding abilities with their paired targets. CONCLUSION: This is a successful case study that validated the application of "seed" mass spectral data could overcome the drawbacks of GNPS libraries with limited mass spectra and enhance the accuracy and visualisation of natural products (NPs) dereplication, and this rapid recognition and targeted isolation strategy can be also applicable for other types of NPs.

Trends in Diet Quality and Increasing Inadequacies of Micronutrients Vitamin C, Vitamin B12, Iron and Potassium in US Type 2 Diabetic Adults.

BACKGROUND: Prevalence of diabetes was high and rose significantly in the US between 1999 and 2018. A healthy dietary pattern that provides micronutrient adequacy is one of the most important lifestyle choices for battling the progress of diabetes. Yet, the patterns and trends of diet quality of the US type 2 diabetes are understudied. OBJECTIVES: We aim to examine the patterns and trends of diet quality and major food sources of macronutrients of US type 2 diabetic adults. METHODS: The 24 h dietary recalls of 7789 type 2 diabetic adults, comprising 94.3% of total adults with diabetes from the US National Health and Nutrition Examination Survey cycles (1999-2018), were analyzed. Diet quality was measured by the total Healthy Eating Index (HEI)-2015 scores and 13 individual components. Trends of usual intakes of vitamin C (VC), vitamin B12 (VB12), iron, and potassium and supplements from two 24 h recalls were also examined for type 2 diabetic population. RESULTS: Diet quality of type 2 diabetic adults worsened between 1999 and 2018 while that of US adults of general population improved based on the total HEI 2015 scores. For people with type 2 diabetes, consumption of saturated fat and added sugar increased while consumption of vegetables and fruits declined significantly, although consumption of refined grain declined and consumption of seafood and plant protein increased significantly. In addition, usual intakes of micronutrients VC, VB12, iron, and potassium from food sources declined significantly during this period. CONCLUSIONS: Diet quality generally worsened for US type 2 diabetic adults between 1999 and 2018. Declining consumptions of fruits, vegetables, and non-poultry meat may have contributed to the increasing inadequacies of VC, VB12, iron, and potassium in the US type 2 diabetic adults.

Genome assembly, resequencing and genome-wide association analyses provide novel insights into the origin, evolution and flower colour variations of flowering cherry.

Flowering cherry is a very popular species around the world. High-quality genome resources for different elite cultivars are needed, and the understanding of their origins and the regulation of key ornamental traits are limited for this tree. Here, a high-quality chromosome-scale genome of Prunus campanulata 'Plena' (PCP), which is a native and elite flowering cherry cultivar in China, was generated. The contig N50 of the genome was 18.31 Mb, and 99.98% of its contigs were anchored to eight chromosomes. Furthermore, a total of 306 accessions of flowering cherry germplasm and six lines of outgroups were collected. Resequencing of these 312 lines was performed, and 761 267 high-quality genomic variants were obtained. The origins of flowering cherry were predicted, and these 306 accessions could be classified into three clades, A, B and C. According to phylogenetic analysis, we predicted two origins of flowering cherry. Flowering cherry in clade A originated in southern China, such as in the Himalayan Mountains, while clades B and C originated in northeastern China. Finally, a genome-wide association study of flower colour was performed for all 312 accessions of flowering cherry germplasm. A total of seven quantitative trait loci (QTLs) were identified. One gene encoding glycosylate transferase was predicted as the candidate gene for one QTL. Taken together, our results provide a valuable genomic resource and novel insights into the origin, evolution and flower colour variations of flowering cherry.

Decreasing Vitamin C Intake, Low Serum Vitamin C Level and Risk for US Adults with Diabetes.

Vitamin C (VC) intakes, serum VC, fasting plasma glucose, and A1c levels of 25,206 adult men and 26,944 adult women with 6807 type 2 and 428 type 1 diabetes from the NHANES database between 1999 and 2018 were analyzed. Our hypothesis is that low VC intake and serum VC level may be a health risk for US adults with diabetes. Analyses revealed total VC intake below the estimated average requirement (EAR) increased from 38.1% to 46.5% between 1999-2018. VC intake and serum VC levels were inversely associated with markers of pre-diabetes and type 2 diabetes, namely, fasting plasma glucose and A1c levels. Risks of type 2 diabetes increased in adults with VC intake below the EAR and with no VC supplement (odds ratio 1.20, 95% CI 1.1-1.3 and 1.28, 95% CI 1.18-1.40, respectively). Median survivor years of diabetic adults with lower and deficient serum VC were shorter than that of diabetic adults with normal serum VC. Mortality risks of type 2 diabetes with low VC intake and/or deficient serum VC levels were elevated compared to those with adequate VC intake and normal serum VC (HR 1.25, 95% CI 1.05-1.49 and 1.84, 95% CI 1.10-3.08, respectively). Observation of declining VC intake and deleterious consequences of low serum VC in US adults with diabetes suggests encouragement of VC intake, including VC supplementation of 500-1000 mg/day, may be beneficial for pre-diabetic and diabetic US adults.

Cell-free chemoenzymatic starch synthesis from carbon dioxide.

Starches, a storage form of carbohydrates, are a major source of calories in the human diet and a primary feedstock for bioindustry. We report a chemical-biochemical hybrid pathway for starch synthesis from carbon dioxide (CO2) and hydrogen in a cell-free system. The artificial starch anabolic pathway (ASAP), consisting of 11 core reactions, was drafted by computational pathway design, established through modular assembly and substitution, and optimized by protein engineering of three bottleneck-associated enzymes. In a chemoenzymatic system with spatial and temporal segregation, ASAP, driven by hydrogen, converts CO2 to starch at a rate of 22 nanomoles of CO2 per minute per milligram of total catalyst, an ~8.5-fold higher rate than starch synthesis in maize. This approach opens the way toward future chemo-biohybrid starch synthesis from CO2.

Non-natural Aldol Reactions Enable the Design and Construction of Novel One-Carbon Assimilation Pathways in vitro.

Methylotrophs utilizes cheap, abundant one-carbon compounds, offering a promising green, sustainable and economical alternative to current sugar-based biomanufacturing. However, natural one-carbon assimilation pathways come with many disadvantages, such as complicated reaction steps, the need for additional energy and/or reducing power, or loss of CO2, resulting in unsatisfactory biomanufacturing performance. Here, we predicted eight simple, novel and carbon-conserving formaldehyde (FALD) assimilation pathways based on the extended metabolic network with non-natural aldol reactions using the comb-flux balance analysis (FBA) algorithm. Three of these pathways were found to be independent of energy/reducing equivalents, and thus chosen for further experimental verification. Then, two novel aldol reactions, condensing D-erythrose 4-phosphate and glycolaldehyde (GALD) into 2R,3R-stereo allose 6-phosphate by DeoC or 2S,3R-stereo altrose 6-phosphate by TalBF178Y/Fsa, were identified for the first time. Finally, a novel FALD assimilation pathway proceeding via allose 6-phosphate, named as the glycolaldehyde-allose 6-phosphate assimilation (GAPA) pathway, was constructed in vitro with a high carbon yield of 94%. This work provides an elegant paradigm for systematic design of one-carbon assimilation pathways based on artificial aldolase (ALS) reactions, which could also be feasibly adapted for the mining of other metabolic pathways.

Ultrastable Zirconium-Based Cationic Metal-Organic Frameworks for Perrhenate Removal from Wastewater.

The effective removal of radioactive 99TcO4- anion from nuclear wastewater remains a very difficult unsolved problem. Functional adsorbent materials with high stability, anion-exchange capacity, excellent selectivity, and recyclability are much needed to solve this problem. In this work, we designed two stable cationic metal-organic frameworks (MOFs)-Zr-tcbp-Me and Zr-tcpp-Me-for possible use as adsorbent materials to remove 99TcO4-. Both compounds were synthesized by solvothermal reactions of the tetracarboxylate ligand with zirconium salt, followed by postsynthetic modification (N-methylation). The crystallinity of both zirconium-based MOFs can be well retained under harsh conditions, and they exhibit high adsorption capacity and selectivity toward ReO4- anion, a nonradioactive analogue of 99TcO4-. Zr-tcbp-Me and Zr-tcpp-Me demonstrate the highest framework stability toward acidity among all previously reported cationic MOFs that have been tested for perrhenate removal from wastewater.

Decreased Iron Intake Parallels Rising Iron Deficiency Anemia and Related Mortality Rates in the US Population.

BACKGROUND: Prevalence of iron deficiency anemia and its related mortality rate are on the rise in the United States and causes are unclear. OBJECTIVES: The aim was to examine trends and causes of Fe deficiency anemia prevalence and Fe deficiency-related mortality rates in the total US population. METHODS: Changes in daily dietary Fe intake, serum iron concentration, hemoglobin, red cell distribution width (RDW), and mean corpuscular volume (MCV) obtained from the laboratory files of NHANES, Fe deficiency anemia-related mortality rates from the CDC, and iron concentrations of US food products from the USDA between 1999 and 2018 were analyzed. RESULTS: Of food items with revised concentrations in USDA Nutrient Database for Standard Reference [SR28 (2015)], 62.4% had lower Fe concentrations than in SR11 (1999). There was a 15.3% reduction in beef (relatively higher in heme iron) and a 21.5% increase in chicken meat consumption in the American diet between 1999 and 2018. Dietary iron intake decreased by ∼6.6% and ∼9.5% for male and female adults, respectively. Increases of prevalence of estimated anemia in the United States ranged from 10.5% to 106% depending on age and sex. Age-adjusted mortality rates with iron deficiency anemia as the underlying cause of death increased from ∼0.04 to ∼0.08 deaths per 100,000 people, whereas all other anemias as the underlying causes of death decreased by ≥25%. Mean RDW and serum folate concentrations increased, whereas hemoglobin, serum iron concentrations, and MCV, parameters traditionally associated with Fe deficiency anemia, decreased during this period. CONCLUSIONS: Increased iron deficiency anemia and related mortality rates in the US population between 1999 and 2018 were likely related to the decline in dietary iron intake resulting from an Fe concentration decline in US food products and a shift in dietary patterns.

Rising Trend of Hypokalemia Prevalence in the US Population and Possible Food Causes.

Background: Potassium intake deficiency is a chronic issue in the US and many other countries. Possible causes of the deficiency are understudied.Objective: This study examined potassium deficiency in the US population and possible causes for the new trend.Methods: Serum potassium data of 28,379 men and 29,617 women between ages 12 and 80 years old who participated in the US National Health and Nutrition Examination Survey (NHANES) between 1999 and 2016 were examined. Blood samples were collected by NHANES and blood biochemistry data were measured in designed laboratories. The data were released bi-annually. Possible causes of low potassium intakes were explored.Results: There was an apparent decline of serum potassium in the US population between ages 12 and 80 years from 1999 to 2016. Annual average serum potassium concentrations changed from 4.14 ± 0.01 to 3.97 ± 0.01 mmol/l during this period. Hypokalemia prevalence in the US rose from 3.78%±0.68% to 11.06%±1.08% during this period with a higher hypokalemia prevalence in non-Hispanic black than in non-Hispanic white persons. It is possible that declining potassium concentration in food sources in the US contributed to lower potassium intake and increasing potassium deficiency.Conclusion: The rising trend of hypokalemia prevalence in the US population between 1999 and 2016 is alarming. Renewed efforts to reduce potassium intake deficiency in the US at population level are needed. The impact of possible decreasing crop available potassium levels and increasing consumption of processed food on the potassium deficit trend in the US are possible explanations for the rise in hypokalemia prevalence and require further study.

Economic burden of maintenance hemodialysis patients' families in Nanchong and its influencing factors.

BACKGROUND: Maintenance hemodialysis is the main therapy for clinical treatment of end-stage renal disease (ESRD). The aim of this study was to analyze the current status of the economic burden levied on families with members who are maintenance hemodialysis patients in Nanchong, and the related influencing factors. METHODS: A total of 111 patients with ESRD who were admitted to our hospital from April 2018 to April 2020 and treated with maintenance hemodialysis were selected as research subjects. A questionnaire survey was adopted as a data collection and interview method to observe the economic burden of families with a member who was a maintenance hemodialysis patient. Logistic regression analysis was used to analyze the independent risk factors that affect this economic burden. RESULTS: The direct economic burden, indirect economic burden, and average annual total economic burden (the sum of the direct economic burden and indirect economic burden of hemodialysis patients) of patients in the resident medical insurance group were significantly higher than those in the employee medical insurance group, resident medical insurance + poverty relief group, and employee medical insurance + poverty relief group (P<0.05). The analysis of the unconditional multifactor logistic regression model showed that age, occupation, monthly family income, and medical insurance type were independent risk factors that affected the average annual total economic burden of patients with maintenance hemodialysis (P<0.01). CONCLUSIONS: Various medical insurance systems can effectively reduce the economic burden of hemodialysis patients, but patients must still bear significant financial hardship. It is necessary to further improve the medical insurance for patients with hemodialysis and increase management efforts to popularize the poverty relief policy.

Rise in Potassium Deficiency in the US Population Linked to Agriculture Practices and Dietary Potassium Deficits.

This paper, for the first time, provides evidence that current practices that lead to agricultural crop removal of potassium are unsustainable and likely contributed to the decline in dietary potassium intake and rise in hypokalemia prevalence in the US population. Potassium concentrations in beef, pork, turkey, fruit, vegetables, cereal crops, and so forth decreased between 1999 and 2015 based on the examination of potassium values of food items of USDA standard reference. Ratios of potassium input to removal by crops between 1987 and 2014, potassium in topsoil, and crop-available soil potassium in US farms all declined in recent years. Reported reductions in dietary potassium intake correspond to these decreases in the food supply and to increases in hypokalemia prevalence in the US population. Results of this paper provide new understanding on links between potassium management in agricultural practices and potassium intake deficits, which is needed for combating increasing hypokalemia prevalence in the US population.

[Comparison of soil hydraulic properties in wild and cultivated areas of Notopterygium incisum].

To clarify the difference of soil moisture characteristics between mixed broad leaf-conifer forest soil and artificial cultivation of Notopterygium incisum, the HYPROP system and the dew point potential meter were used to determine soil water retention curves(SWRC) for samples of two horizons(i.e. 2-7 cm, 10-15 cm). The basic physical and chemical properties of soil and its water characteristic parameters were also determined. The result showed as fllows:①The bulk density of mixed coniferous-broad leaf forest soil was between 0.33 and 0.52 g·cm~(-3), significantly lower than the corresponding value of field soil(1.01-1.18 g·cm~(-3))(P<0.05), While the organic matter content was significantly higher than the corresponding value of field soil(P<0.05). ②The saturated water content(θ_s), field water holding capacity(θ_(FC)) and Water that can be effectively utilized by plants(θ_(PAC)) of mixed coniferous-broadleaved forest soil were significantly higher than the corresponding value of field soil(P<0.05), while the retained water content(θ_r) value that cannot be effectively utilized by plants was significantly lower than that of field soil(P<0.05). ③The values of structural porosity(0.13-0.24 cm~3·cm~(-3)) and Matrix porosity(0.34-0.44 cm~3·cm~(-3)) of mixed coniferous-broadleaved forest soil were higher than the corresponding values of field soil. Therefore, with low bulk density and high content of organic matter, mixed coniferous-broadleaved forest soil can store more water in soil in the form of effective water to meet the needs of plants for water, thus possibly forming high quality medicinal materials of Notopterygii Rhizoma et Radix. In conclusion, the results of this study can provide theoretical basis guidance for soil structure improvement and water management to form high quality medicinal materials in the artificial cultivation of N. incisum.

Exposure to organophosphorus insecticides and increased risks of health and cancer in US women.

Results of this paper provide evidence that chronic long-term exposure to organophosphorus insecticides poses a significantly higher health risk for US women than for men, based on dialkylphosphate biomarker data from NHANES cycles 2003-2012. The risk of cardiovascular disease for female non-smokers aged 60-85 years in the highest dimethylthiophosphate (DMTP) urinary concentration quartile is 3.0 (odds ratio, OD = 3.0, 95%CI 1.4-6.4) times higher than that in the lowest quartile. Women with higher urinary DMTP concentrations also have significantly higher risk of asthma at the ages 6-39 years and an apparently higher risk of chronic bronchitis at the ages 60-85. Overall cancer risk is significantly higher for female non-smokers aged 60-85 years in the higher urinary DMTP quartiles (OD = 2.7, 95% CI 1.3-5.9). Increasing risks of breast cancer for female smokers and prostate cancer for male smokers aged 60-85 years with higher exposure to organophosphorus insecticides in the US are also significant.

[Correlation between growth characteristics and quality of rhizomes of Notopterygium incisum under wild tending].

Study the growth and development process of rhizomes(bamboo-like part) of Notopterygium incisum and the changes of carbohydrate, endogenous hormones and secondary metabolites, and provide theoretical guidance for the formation of high-quality N. incisum medicinal commodities under artificial cultivation. The One-year-old seedlings were transplanted to the original habitat,and the growth and physiological characteristics of N. incisum were dynamically monitored. The results showed that: ① Seedlings transplanted to the original habitat in spring could form rhizomes(bamboo-like part) in the same year. ② After 60 days of transplantation, the root length and root diameter of underground part of N. incisum had increased rapidly, and carbohydrate content in roots and rhizomes had accumulated rapidly. After 120 days of transplantation, the roots and rhizomes of underground part had grown slowly, and starch content in roots and rhizomes increased continuously, while sucrose and total soluble sugar content decreased gradually. ③ The content of abscisic acid(ABA) in rhizomes decreased firstly and then increased, while the indole acetic acid(IAA) content stabilized firstly and then increased rapidly, and the contents of gibberellin(GA_3) and zeatin riboside(ZR) continued to increase. ④ The content of notopterol in rhizomes was higher than that in roots, while the content of isoimperatorin was lower than that in roots, but the total content of the both in rhizomes was higher than that in roots. Therefore, N. incisum can form rhizomes with high content of secondary metabolites under wild tending, and the growth and development of rhizomes are closely related to changes in carbohydrates and are regulated by related endogenous hormones.

[Ethnobotanical study of Juenang cultural area in Rangtang county of northwestern Sichuan].

Indigenous knowledge and traditional culture for sustainable use of native plants in Juenang cultural region of Rangtang county, Aba Zang and Qiang Prefecture of Sichuan province, have been characterized in this paper followed the principles and methods of ethnobotany. The results indicate that 38 species from 27 families(including 6 species of fungi) are ethnobotanically used commonly in this area. Of 38 species of the native plants, 13 species from 12 families are collected for eatables and vegetables, 12 families and 16 species of indigenous plants for medicinal and edible use, 4 species from 4 families for decoration, 4 species from 4 families used for building materials or firewood, and 1 species from 1 families used for religious folklore. Under the influence of Juenang culture and Tibetan culture, indigenous knowledge such as instinctive reverence and gratitude for nature, protection ecological environment and habitats, and moderate use of natural resources(especially wild bioresource), have been gotten passed on from generation to generation in Juenang culture region of Rangtang county, which is of great significance to the protection of local bioresources and environment, including ethnic medicinal plants, and also to provide practical guidance for biodiversity conservation and ecological restoration in those alpine ecological vulnerable areas.